IgG Detection Using Anti- IgG Conjugated Microparticles
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IgG Detection Using IgG Detection Using Anti-IgG Conjugated Anti-IgG Conjugated
MicroparticlesMicroparticles
By Sarah Leung and By Sarah Leung and
Anbar NajamAnbar Najam
MethodsMethods
Conjugated anti-IgG antibodies to 0.92 Conjugated anti-IgG antibodies to 0.92 um microparticles um microparticles
33% microparticle surface coverage with 33% microparticle surface coverage with IgGIgG
0.02% microparticle concentration in 0.02% microparticle concentration in solutionsolution
MethodsMethods
Positioned a 180 degree optical Positioned a 180 degree optical reflection probe directly above reflection probe directly above samples for detection on a two-well samples for detection on a two-well slideslide
ResultsResults 5*105*10-11 -11 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions
12600
12700
12800
12900
13000
13100
13200
13300
13400
13500
13600
13700
1E-12 1E-10 1E-08 0.000001 0.0001 0.01 1
Concentration (g/mL)
Inte
nsi
ty
ResultsResults5*105*10-15 -15 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions
13400.00
13600.00
13800.00
14000.00
14200.00
14400.00
14600.00
14800.00
15000.00
15200.00
1E-17 1E-14 1E-11 0.00000001 0.00001 0.01 10
Concentration (g/mL)
Inte
ns
ity
DiscussionDiscussion
Results lacked evidence of any trends Results lacked evidence of any trends between antigen concentration and light between antigen concentration and light scattering intensityscattering intensity
Possible explanations:Possible explanations: Coagulation of microparticlesCoagulation of microparticles High scattering due to 180 degree detectionHigh scattering due to 180 degree detection Conjugation of antibody, rather than antigen, Conjugation of antibody, rather than antigen,
to microparticlesto microparticles
Next ExperimentNext Experiment
Added TWEEN 80 to existing 0.92 um Added TWEEN 80 to existing 0.92 um microparticle solution and evaluate 180 microparticle solution and evaluate 180 degree vs. 45 degree light scattering detectiondegree vs. 45 degree light scattering detection
180 degrees 45 degreesVs.
ResultsResults
5*105*10-14 -14 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions
ResultsResults
5*105*10-14 -14 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions
DiscussionDiscussion
180 degree results lacked evidence of 180 degree results lacked evidence of any trends between antigen concentration any trends between antigen concentration and light scattering intensityand light scattering intensity
With 45 degree probe, we found a With 45 degree probe, we found a detection limit of 10detection limit of 10-6 -6 g/mlg/ml
Contact Angle Measurements of Contact Angle Measurements of Anti-E. Coli and Anti-E. Coli Anti-E. Coli and Anti-E. Coli
Conjugated Microparticles on Conjugated Microparticles on Superhydrophobic SurfaceSuperhydrophobic Surface
MethodsMethods
Conjugated anti-E. coli antibodies to 0.92 um Conjugated anti-E. coli antibodies to 0.92 um microparticles microparticles
33% microparticle surface coverage with 33% microparticle surface coverage with anti-E. coli anti-E. coli
0.02% microparticle concentration in solution0.02% microparticle concentration in solution
MethodsMethods
Dispensed 10 ul droplet on to superhydrophobic Dispensed 10 ul droplet on to superhydrophobic surface.surface.
Measured contact angle over 10 minutesMeasured contact angle over 10 minutes
Droplets:Droplets: Deionized WaterDeionized Water Anti-E. coli -Conc. 5*10Anti-E. coli -Conc. 5*10-7-7 Anti-E. coli conjugated to microparticlesAnti-E. coli conjugated to microparticles
ResultsResults
Anti-E. coli -Conc. 5*10Anti-E. coli -Conc. 5*10-7-7
DiscussionDiscussion
Results oscillate and do not form a Results oscillate and do not form a smooth curve. smooth curve.
Possible explanations:Possible explanations: Surface contaminationSurface contamination Droplet movement, vibration.Droplet movement, vibration. Position of droplet.Position of droplet. Size of dropletSize of droplet
Next ExperimentNext Experiment
Decrease the volume of the droplet from Decrease the volume of the droplet from 10 ul to 3 ul for a more spherical shape.10 ul to 3 ul for a more spherical shape.
Methods
ResultsResults
Deionized WaterDeionized Water
Future WorkFuture Work
Minimize the oscillations.Minimize the oscillations.
Change the position of the surface.Change the position of the surface.
Determine an efficient way to dispense 3ul Determine an efficient way to dispense 3ul droplet on to superhydrophobic surfacedroplet on to superhydrophobic surface
Acknowledgement s
UA/NASA Space Grant
Dr. Jeong-Yeol Yoon
Lab group: Tremaine B. Powell, Jin-Hee Han, Brian C. Heinze, Phat L. Tran, Jennine N. Chesler, Dr. Keesung
Kim, and Dr. Song
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